U.S. patent number 4,851,536 [Application Number 07/183,861] was granted by the patent office on 1989-07-25 for cyclohexylquinolines as inhibitors of interleukin 1.
This patent grant is currently assigned to American Home Products Corporation. Invention is credited to Steven C. Gilman, Jerauld S. Skotnicki.
United States Patent |
4,851,536 |
Skotnicki , et al. |
July 25, 1989 |
Cyclohexylquinolines as inhibitors of interleukin 1
Abstract
There are disclosed compounds of the formula ##STR1## wherein X
is O, S, SO, SO.sub.2 or CR.sup.1 R.sup.2 ; R.sup.1 and R.sup.2 are
each independently hydrogen, lower alkyl, carboxyl, lower alkoxy
carbonyl, lower cycloalkyl, phenyl, naphthyl, pyridyl, quinolinyl,
pyrazinyl, pyridinyl, pyridazinyl, pyrimidinyl, quinoxalinyl,
quinazolinyl or any of the foregoing aryl or hetaryl substituents
substituted with halo, lower alkyl, lower alkyl carbonyl, benzoyl,
COOR.sup.3, OR.sup.3, N(R.sup.3).sub.2, CON(R.sup.3).sub.2,
SO.sub.3 R.sup.3, SO.sub.2 N(R.sup.3).sub.2, phenylsulfonyl, lower
alkyl sulfonyl, cyano, nitro or trifluoromethyl; R.sup.3 is
hydrogen, lower alkyl or phenyl; R.sup.4 is halo, morpholino,
4-methylpiperazino, R.sup.5 NNHR.sup.6, R.sup.5 NCH.sub.2 CH.sub.2
OCH.sub.3, or ##STR2## R.sup.5 is hydrogen or lower alkyl; R.sup.6
is hydrogen, lower alkyl, lower alkanoyl, lower cycloalkyl or
phenyl; and R.sup.7 and R.sup.8 are each independently, hydrogen,
halo, nitro, lower alkoxy, lower alkyl, cyano, trifluoromethyl,
phenyl, carboxy or lower alkoxycarbonyl, with the proviso that when
R.sup.1 and R.sup.2 are hydrogen or lower alkyl, R.sup.4 is other
than halo. and, by virtue of their ability to inhibit interleukin
1, their use as antinflammatory agents and in treatment of disease
states involving enzymatic tissue destruction.
Inventors: |
Skotnicki; Jerauld S. (Chadds
Ford, PA), Gilman; Steven C. (Berwyn, PA) |
Assignee: |
American Home Products
Corporation (New York, NY)
|
Family
ID: |
26724949 |
Appl.
No.: |
07/183,861 |
Filed: |
April 20, 1988 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
Issue Date |
|
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47376 |
May 7, 1987 |
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Current U.S.
Class: |
546/106; 544/126;
544/284; 544/353; 544/361; 544/405; 546/80; 546/89; 546/102;
546/104; 546/105; 546/107 |
Current CPC
Class: |
C07D
219/04 (20130101); C07D 219/10 (20130101); C07D
495/04 (20130101) |
Current International
Class: |
C07D
219/10 (20060101); C07D 219/04 (20060101); C07D
219/00 (20060101); C07D 495/00 (20060101); C07D
495/04 (20060101); C07D 219/10 (); A61K
031/47 () |
Field of
Search: |
;546/104,102,103,105,106
;544/126,361 |
References Cited
[Referenced By]
U.S. Patent Documents
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3232945 |
February 1966 |
Sigaln, Jr. et al. |
4550113 |
October 1985 |
Lavretskaya et al. |
4608383 |
August 1986 |
Wiedemann et al. |
4631286 |
December 1986 |
Shutake et al. |
4695573 |
September 1987 |
Shutske et al. |
4751305 |
June 1988 |
Skotnicki et al. |
4753950 |
June 1988 |
Shutske et al. |
|
Foreign Patent Documents
Other References
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Abstract of Indian J. Chem., Sect. B 1987, 26B(4), pp. 319-329
(Eng.). .
Brian, et al., Chemical Abstracts, vol. 62: 6459e (1965). .
Konshin, et al., Khin. Fermetsev. Zhur., vol. 5(11), pp. 10-12
(1971) English translation. .
Konshin, et al., Khim. Fermetsev. Zhur., vol. 8(7), pp. 17-19
(1974) English translation. .
Joshi, et al., Indian J. Chem., vol. 16B, pp. 156-158 (02/78).
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Pellerano, et al., Chemical Abstracts, vol. 101: 72587y (1984).
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Patraik, et al., J. Med. Chem., vol. 9, pp. 483-488 (07/66). .
Steinberg, et al., J. Med. Chem., vol. 18(11), pp. 1056-1061
(1975). .
Bielavsky, Coll. Czech. Chem. Commun., vol. 42, pp. 2802-2808
(1977). .
Sargent, et al., Chemical Abstracts, vol. 41 442b-h (1947). .
Sargent, et al., (II), Chemical Abstracts, vol. 41: 7398a-f (1947).
.
Artico, et al., Chemical Abstracts, vol. 64: 9683b-e (1966). .
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Nasr, et al., Chemical Abstracts, vol. 89: 163459h (1978)..
|
Primary Examiner: Daus; Donald G.
Assistant Examiner: Rivers; Diana G.
Attorney, Agent or Firm: Tarnowski; George
Parent Case Text
This is a continuation-in-part of U.S. Ser. No. 47,376, filed May
7, 1987 now abandoned.
Claims
What is claimed is:
1. A compound having the formula ##STR10## wherein R.sup.1 and
R.sup.2 are each independently hydrogen, lower alkyl, carboxy,
lower alkoxy carbonyl, lower cycloalkyl, phenyl, naphthyl, pyridyl,
quinolinyl, or any of the foregoing aryl or hetaryl substituents
substituted with halo, lower alkyl, COOR.sup.3, OR.sup.3,
N(R.sup.3).sub.2, CON(R.sup.3).sub.2, phenylsulfonyl, lower alkyl
sulfonyl, cyano, nitro or trifluoromethyl;
R.sup.3 is hydrogen, lower alkyl or phenyl;
R.sup.4 is halo, or R.sup.5 NNHR.sup.6
R.sup.5 is hydrogen or lower alkyl;
R.sup.6 is lower alkyl, lower alkanoyl, lower cycloalkyl or phenyl;
and
R.sup.7 and R.sup.8 are each independently, hydrogen, halo, nitro,
lower alkoxy, lower alkyl, cyano, trifluoromethyl, phenyl, carboxy
or lower alkoxycarbonyl, with the proviso that when R.sup.1 and
R.sup.2 are hydrogen or lower alkyl, R.sup.4 is other than
halo.
2. The compound of claim 1, having the name
6-chloro-1,2,3,4-tetrahydro-2-methyl-9-(2-phenylhydrazino)acridine.
3. The compound of claim 1, having the name
6-chloro-1,2,3,4-tetrahydro-2-phenyl-9-(2-phenylhydrazino)acridine.
4. The compound of claim 1, having the name
6,9-dichloro-1,2,3,4-tetrahydro-2-acridinecarboxylic acid methyl
ester.
5. The compound of claim 1, having the name
6-chloro-1,2,3,4-tetrahydro-9-(2-phenylhydrazino)-2-acridinecarboxylic
acid ethyl ester.
6. The compound of claim 1, having the name
6,9-dichloro-1,2,3,4-tetrahydro-2-acridinecarboxylic acid.
7. The compound of claim 1, having the name
6-chloro-1,2,3,4-tetrahydro-9-(2-phenylhydrazino)-2-acridinecarboxylic
acid.
8. The compound of claim 1, having the name
6,9-dichloro-1,2,3,4-tetrahydro-2-phenylacridine.
Description
This invention relates to novel compounds possessing interleukin 1
(IL 1) antagonist activity and having antiinflammatory
activity.
Interleukin 1 (IL 1) is a peptide hormone exhibiting a number of
immune and inflammatory actions [Dinarello, Rev. Inf. Dis. 6, 51
(1984)]. IL 1 is produced, in response to inflammatory stimuli, by
leukocytes such as macrophages and polymorphonuclear cells, as well
as by a variety of other cell types such as synovial cells,
endothelial cells and keratinocytes, and it mediates several
biological responses of leukocytes on other tissue targets such as
bone, articular joints, liver, hypothalamus, and brain.
IL 1 was originally shown to augment the proliferation of T
lymphocytes for which it was named lymphocyte activating factor
(LAF), and is believed to be important for the generation of T
cell-dependent immune responses.
There is evidence to suggest a relationship between IL 1 and
pathology in various diseases, particularly immunoinflammatory
disorders such as rheumatoidarthritis [Dinarello et al., Ann. Rev.
Med. 37, 173 (1986)]. IL 1 induces acute inflammatory responses
producing soft tissue swelling (edema and erythema) [Granstein et
al., J. Clin. Invest., 77, 1010 (1986)]. It is a chemoattractant
for polymorphonuclear leukocytes (PMN) and induces the activation
and migration of these cells into tissues. IL 1 also stimulates the
production of prostaglandin E.sub.2, a potent inflammatory
arachidonic acid metabolite, by a variety of cells and tissues
including chondrocytes and synovial cells [Mizel et al., Proc.
Nat'l Acad. Sci., 78, 2474 (1981) and Chang et al., J. Immunol.,
136, 1283 (1986)] and hypothalamic tissue. This effect on the
hypothalamus is thought to be responsible for fever production. IL
1 can induce articular joint destruction by stimulating the
production of a variety of hydrolytic enzymes (neutral proteases
such as collagenase, glycosaminoglycanases, etc.) which degrade
cartilage matrix proteins (collagen, proteoglycan, etc.) by
synovial cells, chondrocytes, and fibroblasts [Dayer et al.,
Science, 195, 181 (1977) and Postlethwaite et al., J. Exp. Med.,
157, 801 (1983)]. Furthermore, IL 1 induces hyperproliferation of
dermal and synovial fibroblasts and is a potent inducer of bone
resorption [Wood et al., J. Immunol., 134, 895 (1985) and Gilman
and Kimball, Agents and Actions, 16, 468 (1985)].
Finally, IL 1 mediates acute phase reactions including alterations
in plasma divalent cations, increased synthesis by liver cells of
acute phase proteins (C-reactive protein, serum amyloid A, etc.)
and fever. Accordingly, compounds which have IL 1 antagonist
activity and thereby inhibit the biological effects of IL 1 can be
advantageously used to block pathologies in which one or more of
these events occur such as rheumatoid arthritis, osteoarthritis and
related disorders [Rodnan and Schumacher, eds, "Primer on the
Arthritic Diseases" 8 ed. Atlanta, 1983], psoriasis and other
inflammatory/proliferative skin disorders as well as diseases in
which the secretion of collagenase (and other tissue hydrolysing
neutral proteinases) has been implicated as a causative factor,
including periodontal disease, tumor invasiveness, and
epidermolysis bullosa [Perez-Tamayo, Amer. J. Pathol., 92, 509
(1978) and Harris and Krane, N. Engl. J. Med., 291, 652 (1974)] and
so forth.
It has now been found that certain novel pyrano-, thiopyrano- and
cyclohexyl-quinolines antagonize the activity of IL 1, and so are
useful as antiinflammatory agents and in the treatment of
pathologies whose etiology is collagenase-based tissue destruction.
The present invention provides novel compounds having the formula:
##STR3## wherein X is O, S, SO, SO.sub.2 or CR.sup.1 R.sup.2 ;
R.sup.1 and R.sup.2 are each independently hydrogen, lower alkyl,
carboxyl, lower alkoxy carbonyl, lower cycloalkyl, phenyl,
naphthyl, pyridyl, quinolinyl, pyrazinyl, pyridinyl, pyridazinyl,
pyrimidinyl, quinoxalinyl, quinazolinyl or any of the foregoing
aryl or hetaryl substituents substituted with halo, lower alkyl,
lower alkyl carbonyl, benzoyl, COOR.sup.3, OR.sup.3,
N(R.sup.3).sub.2, CON(R.sup.3).sub.2, SO.sub.3 R.sup.3, SO.sub.2
N(R.sup.3).sub.2, phenylsulfonyl, lower alkyl sulfonyl, cyano,
nitro or trifluoromethyl;
R.sup.3 is hydrogen, lower alkyl or phenyl;
R.sup.4 is halo, morpholino, 4-methylpiperazino, R.sup.5
NNHR.sup.6,
R.sup.5 NCH.sub.2 CH.sub.2 OCH.sub.3 or ##STR4## R.sup.5 is
hydrogen or lower alkyl; R.sup.6 is hydrogen, lower alkyl, lower
alkanoyl, lower cycloalkyl or phenyl; and
R.sup.7 and R.sup.8 are each independently, hydrogen, halo, nitro,
lower alkoxy, lower alkyl, cyano, trifluoromethyl, phenyl, carboxy
or lower alkoxycarbonyl, with the proviso that when R.sup.1 and
R.sup.2 are hydrogen or lower alkyl, R.sup.4 is other than
halo.
The terms "lower alkyl" and "lower alkoxy" refer to moieties having
1 to 6 carbon atoms in the carbon chain. The term "lower alkanoyl"
refers to the moiety RCO- wherein R is an alkyl group having 1 to 6
carbon atoms. The term "lower cycloalkyl" refers to a saturated
ring having 4 to 7 carbon atoms. The term "halo" refers to fluoro,
chloro and bromo.
The especially preferred compounds are those having the formula
##STR5## wherein X is S, SO.sub.2, CHCH.sub.3 or ##STR6## and
R.sup.4 is chloro or ##STR7##
The compounds of the invention can be prepared by the reaction of a
suitable cyclohexanone, tetrahydropyran-4-one or
tetrahydrothiopyran-4-one with a suitably substituted amino benzoic
acid in the presence of a halogenating agent to yield a halogenated
intermediate: ##STR8## followed by reaction of the intermediate so
obtained with a suitably substituted R.sup.4 -containing reactant
to yield the desired final product: ##STR9##
The starting materials used in the above outlined preparative
sequences are all available commercially or can be prepared by
conventional methods disclosed in the chemical literature.
The compounds of the invention, by virtue of the ability to
antagonize interleukin 1, are useful in the treatment of such
diseases as rheumatoid arthritis, osteoarthritis, tendinitis,
bursitis and similar conditions involving inflammation, as well as
psoriasis and other inflammatory/proliferative skin disorders.
Moreover, the compounds are useful in treating disease states
involving enzymatic tissue destruction, for example, conditions in
which collagenase has been implicated as a causative factor, such
as rheumatoid arthritis joint destruction, periodontal disease,
tumor invasiveness, corneal ulcerations, epidermolysis bullosa and
the like.
When the compounds of the invention are employed as
antiinflammatory agents, or collagenase inhibitors, they can be
formulated into oral dosage forms such as tablets, capsules and the
like. The compounds can be administered alone or by combining them
with conventional carriers, such as magnesium carbonate, magnesium
stearate, talc, sugar, lactose, pectin, dextrin, starch, gelatin,
tragacanth, methylcellulose, sodium carboxymethylcellulose, low
melting wax, cocoa butter and the like. Diluents, flavoring agents,
solubilizers, lubricants, suspending agents, binders,
tablet-disintegrating agents and the like may be employed. The
compounds may be encapsulated with or without other carriers. In
all cases, the proportion of active ingredients in said
compositions both solid and liquid will be at least to impart the
desired activity thereto on oral administration. The compounds may
also be injected parenterally, in which case they are used in the
form of a sterile solution containing other solutes, for example,
enough saline or glucose to make the solution isotonic. For topical
administration, the compounds may be formulated in the form of
dusting powders, solutions, creams, lotions or aerosols in
pharmaceutically acceptable vehicles, which are applied to affected
portions of the skin.
The dosage requirements vary with the particular compositions
employed, the route of administration, the severity of the symptoms
presented and the particular subject being treated. Treatment will
generally be initiated with small dosages less than the optimum
dose of the compound. Thereafter the dosage is increased until the
optimum effect under the circumstances is reached. In general, the
compounds of the invention are most desirably administered at a
concentration that will generally afford effective results without
causing any harmful or deleterious side effects, and can be
administered either as a single unit dose, or if desired, the
dosage may be divided into convenient subunits administered at
suitable times throughout the day.
The interleukin 1 antagonist activity of the compounds of the
invention may be demonstrated by standard pharmacological
procedures, which are described more fully in the examples given
hereinafter.
These procedures illustrate the ability of the compounds of the
invention to inhibit the IL 1-induced release of neutral protease
from articular chondrocytes.
The following examples show the preparation and pharmacological
testing of compounds within the invention.
EXAMPLE 1
7-Chloro-3,4-dihydro-10-(2-phenylhydrazino)-1H-thiopyrano[4,3-b]quinoline
A. 7,10-Dichloro-3,4-dihydro-1H-thiopyrano[4,3-b]quinoline
To a slurry of 14.8 g (0.0863 mol) of 2-amino-4-chlorobenzoic acid
and 150 ml of phosphorous oxychloride is added dropwise 10 g (0.086
mol) of tetrahydrothiopyran-4-one. The mixture is stirred at reflux
for 3 hours and then concentrated in vacuo. The residue is
dissolved in methylene chloride and edded slowly to an ice-NH.sub.4
OH mixture. The mixture is stirred for 1/2 hour and extracted with
methylene chloride. The combined extracts are washed with water,
dried over Na.sub.2 SO.sub.4, and concentrated in vacuo to yield a
dark solid. Trituration with ether furnishes 15 g (64%) of title
compound: m.p. 112.degree.-115.degree. C.; IR (KBr) 1605, 1480, and
1415 cm.sup.-1 ; NMR (CDCl.sub.3) .delta. 8.08-8.02 (m, 2H),
7.58-7.52 (m, 1H), B 4.08 (s, 2H), 3.44 (t, 2H), and 3.09 (t,
2H).
Analysis for: C.sub.12 H.sub.9 NCl.sub.2 S Calculated: C, 53.34; H,
3.36; N, 5.19. Found: C, 52.98; H, 3.51; N, 5.61.
B.
7-Chloro-3,4-dihydro-10-(2-phenylhydrazino)-1H-thiopyrano[4,3-b]quinoline
A mixture of 4.0 g (0.015 mol) of the compound of step A, above,
3.2 ml (0.03 mol) of phenylhydrazine, 2.5 ml of concentrated
hydrochloric acid, and 150 ml of absolute ethanol is stirred under
reflux for 6 hours. The precipitate, on cooling, is collected and
dissolved in methanol. Treatment with a Na.sub.2 CO.sub.3 solution
yields an off-white solid. Recrystallization from toluene/hexane
furnishes 575 mg (11%) of title compound: m.p.
201.degree.-202.degree. C.; IR 3300, 3240, 1605, and 1560 cm.sup.-1
; NMR (DMSO-d.sub.6) .delta. 8.68-8.58 (m, 1H), 8.50 (s, 1H,
exchangeable), 8.28 (s, 1H, exchangeable), 7.86-7.8 (m, 1H),
7.46-7.38 (m, 1H), 7.24-6.94 (m, 2H), 6.86-6.72 (m, 3H), 4.04 (s,
2H), 2.20 (t, 2H), and 1.96 (t, 2H).
Analysis for: C.sub.18 H.sub.16 N.sub.3 ClS Calculated: C, 63.24;
H, 4.72; N, 12.29. Found: C, 63.59; H, 4.82; N, 11.93.
EXAMPLE 2
6-Chloro-1,2,3,4-tetrahydro-2-methyl-9-(2-phenylhydrazino)acridine
A. 6,9-Dichloro-1,2,3,4-tetrahydro-2-methylacridine, quarter
hydrate
To a slurry of 15.3 g (0.0892 mol) of 2-amino-4-chlorobenzoic acid
and 150 ml of phosphorous oxychloride is added dropwise 10 g
(0.0892 mol) of 4-methylcyclohexanone. The mixture is stirred at
reflux for 3 hours and then concentrated in vacuo. The residue is
dissolved in methylene chloride and added slowly to an ice-NH.sub.4
OH mixture. The mixture is stirred for 1/2 hour and extracted with
methylene chloride. The combined extracts are washed with water,
dried over Na.sub.2 SO.sub.4, and concentrated in vacuo to yield a
dark solid. Trituration with ether affords 14.2 g (60%) of title
compound: m.p. 85.degree.-87.degree. C.; IR (KBr) 2920, 1605, 1585,
1480, and 1420 cm.sup.-1 ; NMR (CDCl.sub.3) .delta. 8.12-8.0 (m,
2H), 7.50-7.42 (m, 1H), 3.18-3.02 (m, 3H), 2.32-2.20 (m, 1H),
2.12-1.92 (m, 2H), 1.66-1.48 (m, 1H), and 1.2 (d, 3H).
Analysis for: C.sub.14 H.sub.13 NCl.sub.2.1/4H.sub.2 O Calculated:
C, 62.12; H, 5.02; N, 5.18. Found: C, 62.41; H, 4.92; N, 5.49.
B.
6-Chloro-1,2,3,4-tetrahydro-2-methyl-9-(2-phenylhydrazino)acridine
A mixture of 4.0 g (0.015 mol) of the compound of step A, above,
3.2 ml (0.03 mol) of phenylhydrazine, 2.5 ml of concentrated
hydrochloric acid, and 150 ml of absolute ethanol is stirred under
reflux for 6 hours. The precipitate, on cooling, is collected and
dissolved in methanol. Treatment with a Na.sub.2 SO.sub.3 solution
yields an off-white solid. Recrystallization from toluene/hexane
furnishes 1.1 g (22%) of title compound: m.p.
197.degree.-198.degree. C.; IR (KBr) 3330, 3240, 1605, 1555 and
1485 cm.sup.-1 ; NMR (DMSO-d.sub.6) .delta. 8.78-8.70 (m, 1H), 8.18
(s, 1H, exchangeable), 8.08 (s, 1H, exchangeable), 7.76-7.70 (m,
1H), 7.28-7.22 (m, 1H), 7.20-7.12 (m, 2H), 6.84-6.70 (m, 3H),
3.06-2.88 (m, 3H), 2.38-2.24 (m, 1H), 1.98-1.78 (m, 2 H), 1.54-1.38
(m, 1H), and 1.08 (d, 3H).
Analysis for: C.sub.20 H.sub.20 N.sub.3 Cl Calculated: C, 71.10; H,
5.97; N, 12.44. Found: C, 71.12; H, 6.10; N, 12.09.
EXAMPLE 3
6-Chloro-1,2,3,4-tetrahydro-2-phenyl-9-(2-phenylhydrazino)acridine
A. 6,9-Dichloro-1,2,3,4-tetrahydro-2-phenylacridine,
hemihydrate
To a slurry of 15 g (0.0874 mol) of 2-amino-4-chlorobenzoic acid
and 150 ml of phosphorous oxychloride is added dropwise 15 g
(0.0871 mol) of 4-phenylcyclohexanone. The mixture is stirred at
reflux for 3 hours and then concentrated in vacuo. The residue is
dissolved in methylene chloride and added slowly to an ice-NH.sub.4
OH mixture. The mixture is stirred for 1/2 hour and extracted with
methylene chloride. The combined extracts are washed with water,
dried over Na.sub.2 SO.sub.4, and concentrated in vacuo to yield a
dark solid. Trituration with ether provides 20 g (70%) of title
compound: m.p. 134.degree.-136.degree. C.; IR 1605, 1585 and 1480
cm.sup.-1 ; NMR (CDCl.sub.3) .delta. 8.12-8.0 (m, 2H), 7.54-7.46
(m, 1H), 7.44-7.26 (m, 5H), 3.50-2.88 (m, 5H), and 2.28-2.02 (m,
2H).
Analysis for: C.sub.19 H.sub.15 NCl.sub.2.1/2H.sub.2 O Calculated:
C, 67.66; H, 4.78; N, 4.15. Found: C, 67.56; H, 4.57; N, 4.58.
B.
6-Chloro-1,2,3,4-tetrahydro-2-phenyl-9-(2-phenylhydrazino)acridine
A mixture of 5.22 g (0.0159 mol) of the compound of step A, above,
3.2 ml (0.03 mol) of phenylhydrazine, 2.5 ml of concentrated
hydrochloric acid, and 150 ml of absolute ethanol is stirred under
reflux for 6 hours. The precipitate on cooling is collected and
dissolved in methanol. Treatment with Na.sub.2 CO.sub.3 solution
yields an off-white solid. Recrystallization from toluene/hexane
furnishes 2.7 g (42%) of title compound: m.p.
198.degree.-199.degree. C.; IR (KBr) 3320, 3240, 1605, 1555 and
1485 cm.sup.-1 ; NMR (DMSO-d.sub.6) .delta. 8.84-8.76 (m, 1H), 8.22
(s, 1H, exchangeable), 8.16 (s, 1H, exchangeable), 7.78-7.72 (m,
1H), 7.46-7.12 (m, 8H), 6.87-6.70 (m, 3H), 3.28-2.74 (m, 5H), and
2.18-1.90 (m, 2H).
Analysis for: C.sub.25 H.sub.22 N.sub.3 Cl Calculated: C, 75.08; H,
5.55; N, 10.51. Found: C, 74.88; H, 5.60; N, 10.74.
EXAMPLE 4
10-Chloro-3,4-dihydro-1H-thiopyrano[4,3-b]quinoline
A slurry of 10 g (0.0807 mol) of tetrahydropyranone, 11.8 g (0.0807
mol) of anthranilic acid, and 150 ml of phosphorous oxychloride is
stirred at 100.degree. C. for 5 hours, cooled, and concentrated in
vacuo. The residue is dissolved in methylene chloride and added
slowly to an ice-NH.sub.4 OH mixture. The aqueous phase is
separated and extracted with methylene chloride. The combined
organic phases are dried over Na.sub.2 SO.sub.4 and concentrated in
vacuo to give a waxy solid. Trituration with ethyl ether furnishes
11.2 g (59%) of the title compound: IR (KBr) 1575, 1550, and 1480
cm.sup.-1 ; NMR (DMSO-d.sub.6) .delta. 8.18-7.66 (m, 4H), 4.04 (s,
2H), 3.30 (t, 2H), and 3.02 (t, 2H).
Analysis for: C.sub.12 H.sub.10 NSCl Calculated: C, 61.14; H, 4.28;
N, 5.94. Found: C, 61.50; H, 4.39; N, 5.93.
EXAMPLE 5
3,4-Dihydro-10-(2-phenylhydrazino)-1H-thiopyrano[4,3-b]quinoline,
three-quarters hydrate
A mixture of 4.0 g (0.017 mol) of the compound of Example 4, 3.65
ml (3.68 g/0.034 mol) of phenylhydrazine, 150 ml of ethanol, and 2
ml of concentrated hydrochloric acid is stirred at reflux for 6
hours, then allowed to cool to ambient temperature. The resulting
precipitate is dissolved in methanol and this solution is treated
with Na.sub.2 CO.sub.3 solution. The resulting precipitate is
collected and triturated with ethyl ether to afford 980 mg (18%) of
the title compound: IR (KBr) 3280, 3200, 1590, 1560, 1525, and 1490
cm.sup.-1 ; NMR (DMSO-d.sub.6) .delta. 8.58-6.7 (complex m, 9H),
8.36 (br-s, 1H, exchangeable), 8.20 (br-s, 1H, exchangeable), 4.08
(s, 2H), 3.18 (t, 2H), and 2.94 (t, 2H).
Analysis for: C.sub.18 H.sub.17 N.sub.3 S.3/4H.sub.2 O Calculated:
C, 67.36; H, 5.81; N, 13.09. Found: C, 67.36; H, 5.42; N,
12.77.
EXAMPLE 6
10-Chloro-3,4-dihydro-1H-thiopyrano[4,3-b]quinoline 2,2-dioxide
To a mixture of 3 g (0.0127 mol) of the compound of Example 4 and
100 ml of chloroform is added dropwise a solution of 6.04 g (0.028
mol) of m-chloroperoxybenzoic acid and 100 ml of chloroform. The
reaction mixture is stirred for one hour, washed with Na.sub.2
CO.sub.3 solution, dried over Na.sub.2 SO.sub.4, and concentrated
in vacuo to give a pasty solid. Trituration with ethyl ether
furnishes 1.1 g (32%) of the title compound; IR (KBr) 1480, 1320,
and 1120 cm.sup.-1 ; NMR (DMSO-d.sub.6) .delta. 8.30-7.6 (complex
m, 4H), 4.75 (s, 2H), 3.4-3.10 (complex m, 4H).
Analysis for: C.sub.12 H.sub.10 NClO.sub.2 S Calculated: C, 53.83;
H, 3.77; N, 5.23. Found: C, 53.90; H, 3.83; N, 5.59.
EXAMPLE 7
7,10-Dichloro-3,4-dihydro-1H-thiopyrano[4,3-b]quinoline
2,2-dioxide
To a mixture of 3 g (0.0111 mol) of the compound of Example 1A,
above, and 100 ml of chloroform is added dropwise a solution of
5.27 g (0.0244 mol) of m-chloroperoxybenzoic acid and 100 ml of
chloroform. The reaction mixture is stirred for one hour, washed
with Na.sub.2 CO.sub.3 solution, dried over Na.sub.2 SO.sub.4 and
concentrated in vacuo to give a pasty solid. Trituration with ethyl
ether affords 2.65 g (79%) of the title compound: IR (KBr) 1600,
1580, 1540, 1470, 1400, 1305, and 1120 cm.sup.-1 ; NMR
(DMSO-d.sub.6) .delta. 8.30-7.60 (m, 3H), 4.75 (s, 2H), and
3.40-3.05 (m, 4H).
Analysis for: C.sub.12 H.sub.9 NCl.sub.2 O.sub.2 S.1/4H.sub.2 O
Calculated: C, 46.99; H, 3.12; N, 4.57. Found: C, 46.79; H, 3.09;
N, 4.46.
EXAMPLE 8
7-Chloro-3,4-dihydro-10-(2-phenylhydrazino)-1H-thiopyrano[4,3-b]quinoline
2,2-dioxide
To a solution of 110 mg (0.00032 mol) of the compound of Example
1B, above, and 25 ml of chloroform is added dropwise a solution of
185 mg (0.00087 mol) of m-chloroperoxybenzoic acid and 25 ml of
chloroform. The reaction mixture is stirred for one hour, washed
with Na.sub.2 CO.sub.3 solution, dried over Na.sub.2 SO.sub.4, and
concentrated in vacuo. Trituration with ethyl ether affords 32 mg
(27%) of the title compound: IR (KBr) 3400 (br), 3080, 3000, 1600,
1310, 1295, and 1130 cm.sup.-1 ; NMR (DMSO-d.sub.6) .delta.
8.18-7.68 (m, 10H), 4.72 (s, 2H), 3.78-3.64 (m, 4H).
Analysis for: C.sub.18 H.sub.16 N.sub.3 ClSO.sub.2 Calculated: C,
57.83; H, 4.31; N, 11.24. Found: C, 58.46; H, 4.02; N, 10.96.
EXAMPLE 9
6,9-Dichloro-1,2,3,4-tetrahydro-2-acridinecarboxylic acid methyl
ester
To a slurry of 16.5 g (0.096 mol) of 4-chloroanthranilic acid and
166 ml of phosphorous oxychloride is added portionwise 15 g (0.096
mol) of methyl 4-ketocyclohexane carboxylate. The reaction mixture
is refluxed for 3 h, cooled, and concentrated in vacuo. The residue
is dissolved in methylene chloride and poured into an ice-NH.sub.4
OH mixture. The aqueous phase is separated and extracted with
methylene chloride. The combined organic phases are dried over
Na.sub.2 SO.sub.4 and concentrated in vacuo. The residue is
triturated with ethyl ether to give 21.7 g (73%) of the title
compound: IR (KBr) 1730, 1605, 1580, and 1550 cm.sup.-1 ; NMR
(DMSO-d.sub.6) .delta. 8.14-8.08 (m, 1H), 8.0-7.96 (m, 1H),
7.70-7.64 (m, 1H), 3.70 (s, 3H), 3.30-3,16 (m, 1H), 3.12-2.96 (m,
4H), 2.32-2.20 (m, 1H), and 2.04-1.86 (m, 1H).
Analysis for: C.sub.15 H.sub.13 Cl.sub.2 NO.sub.2 Calculated: C,
58.08; H, 4.22; N, 4.52. Found: C, 57.88; H, 4.41; N, 4.59.
EXAMPLE 10
6-Chloro-1,2,3,4-tetrahydro-9-(2-phenylhydrazino)-2-acridinecarboxylic
acid ethyl ester
A mixture of 11 g (0.035 mol) of the compound of Example 9, 7.1 g
(0.066 mol) of phenylhydrazine, 5.5 ml of conc. hydrochloric acid,
and 300 ml of ethanol is heated at reflux overnight. On cooling,
the resulting precipitate is collected, and dissolved in methanol.
This solution is treated with saturated Na.sub.2 CO.sub.3 solution.
The resulting solid is collected, washed with water and triturated
with methylene chloride-ethyl acetate to give 5.7 g (41%) solid.
Recrystallization from isopropanol affords 794 mg (6%) of the title
compound: mp 181.degree.-182.degree. C.; IR (KBr) 3320, 3240, 1720,
1605, and 1550 cm.sup.-1 ; NMR (DMSO-d.sub.6) .delta. 8.24 (br-s,
1H, exchangeable), 8.18 (br-s, 1H, exchangeable), 7.32-7.22 (m,
1H), 7.20-7.16 (m, 3H), 6.86-6.68 (m, 4H), 4.10 (q, 2H), 3.16-2.80
(m, 5H), 2.20-2.06 (m, 1H), 1.96-1.84 (m, 1H), and 1.16 (t,
3H).
Analysis for: C.sub.22 H.sub.22 ClN.sub.3 O.sub.2 Calculated: C,
66.74; H, 5.60; N, 10.62. Found: C, 66.83; H, 5.48; N, 10.49.
EXAMPLE 11
6,9-Dichloro-1,2,3,4-tetrahydro-2-acridinecarboxylic acid
A mixture of 9.9 g (0.032 mol) of the compound of Example 9, 3.25
ml of ethanol, 32.5 ml of water, and 2.6 g of NaOH is stirred
overnight at ambient temperature. The resulting solid is collected,
washed with ethyl acetate, then dissolved in 100 ml of water. The
solution is acidified with acetic acid and the resulting
precipitate is collected, washed copiously with water and dried to
give 6.64 g (70%) of the title compound: mp >250.degree. C.; IR
(KBr) 2900 (br), 2500 (br), 1700 (br), 1600, 1540, and 1470
cm.sup.-1 ; NMR (DMSO-d.sub.6) .delta. 12.6 (br-s, 1H,
exchangeable), 7.92-7.86 (m, 1H), 7.82-7.76 (m, 1H), 7.54-7.48 (m,
1H), 3.10-2.76 (m, 5H), 2.28-2.12 (m, 1H), and 1.94-1.88 (m,
1H).
Analysis for: C.sub.14 H.sub.11 Cl.sub.2 NO.sub.2 Calculated: C,
56.78; H, 3.74; N, 4.73. Found: C, 56.85; H, 3.92; N, 4.82.
EXAMPLE 12
6-Chloro-1,2,3,4-tetrahydro-9-(2-phenylhydrazino)-2-acridinecarboxylic
acid hemihydrate
A mixture of 2.0 g (0.0051 mol) of the compound of Example 10, 6.4
ml of water, 6.4 ml of ethanol, and 0.4 g of NaOH is stirred at
ambient temperature overnight, then concentrated in vacuo. The
residue is dissolved in water and extracted with ethyl acetate. The
aqueous phase is acidified using acetic acid and extracted with
methylene chloride. The organic extracts are dried over Na.sub.2
SO.sub.4, and concentrated in vacuo. The resulting powder is
triturated with petroleum ether to give 42 mg of the title
compound: IR (KBr) 3250 (br), 2940 (br), 2500 (br), 1700, and 1600
cm.sup.-1 ; NMR (DMSO-d.sub.6) .delta. 8.18-7.58 (m, 11H),
3.24-2.90 (m, 5H), 2.38-2.22 (m, 1H), and 2.10-1.98 (m, 1H).
Analysis for: C.sub.20 H.sub.18 ClN.sub.3 O.sub.2.1/2H.sub.2 O
Calculated: C, 63.74; H, 5.08; N, 11.15. Found: C, 63.57; H, 4.55;
N, 10.59.
EXAMPLE 13
The ability of the compounds of the inventions to inhibit
interleukin 1 is measured by the ability of the test compounds to
inhibit the IL 1-induced release of neutral protease from rabbit
articular chondrocytes.
This assay is carried out as follows:
Isolation of rabbit chondrocytes:
Male New Zealand White rabbits are anesthetized with 50 mg/kg of
ketamine (i.m.) and killed by an intracardiac injection of 3 mls of
Nembutal. The knee joints of both legs are resected and the
articular surfaces are exposed. Cartilage slices are obtained using
a scalpel and are placed in a tissue culture dish (100 mm diameter)
containing 10 mls of Hank's balanced salt solution (HBSS). The
chondrocytes within the cartilage slices are then liberated by a
series of enzyme digestions. The slices are incubated for 10
minutes at 37.degree. C. in 0.05% hyaluronidase (Sigma H-3884),
rinsed with HBSS and incubated with 0.2% trypsin (Sigma T-2395) for
10 minutes at 37.degree. C. The slices are rinsed again and
incubated for 10 minutes at 37.degree. C. with 1.2% collagenase
(Sigma C-5138). The slices are then rinsed again with HBSS and
resuspended in 10 ml of Ham's F-12 medium containing 10% fetal
bovine calf serum (FCS) and 0.2% collagenase and incubated
overnight at 37.degree. C. in a 5% CO.sub.2 incubator. The next
day, the medium containing the digested cartilage fragments and
liberated chondrocytes is transferred to a 15 ml centrifuge tube
and the cells are collected by centrifugation and washed twice and
resuspended in Ham's F-12 medium. The cells are then plated into
24-well tissue culture plates (2.times.10.sup.5 cells/well) and
incubated at 37.degree. C. until confluent (usually 4-6 days).
Stimulation of chondrocytes and drug treatment
The confluent chondrocytes are rinsed twice with serum-free Ham's
F-12 medium and 1 ml is added to each well. Fifty .mu.l of purified
human IL 1 (100 Units/ml; Genzyme Corporation, Boston, MA) is then
added to stimulate these cells to secrete neutral protease. To
measure drug effects, the cells are treated with test compound 10
minutes prior to additon of IL 1. The standard screening dose is 10
.mu.M. Twenty-four hours after IL 1 stimulation, supernatant fluids
are collected and assayed for neutral protease activity.
Neutral protease assay
The neutral protease activity of chondrocyte supernatant fluids is
determined by their ability to degrade an insoluble protease
substrate, azocoll (Sigma). Supernatants are treated for 10 minutes
at room temperature with 350 .mu.M p-aminophenylmurcuric acetate to
activate the latent enzyme. Three hundred .mu.l of supernatant is
then mixed with 500 .mu.l of a 20 mg/ml suspension of azocoll and
incubated at 37.degree. C. for 18-24 hours with gentle rocking. The
mixtures are centrifuged and the amount of substrate hydrolyzed is
determined by measuring the absorbance of the supernatant at 520
nm.
Drug effects are calculated as the % change in enzyme activity
(absorbance) by supernatants from drug-treated chondrocytes
relative to enzyme activity of supernatants from vehicle-treated
chondrocytes as follows:
% Inhibition of Protease Secretion= ##EQU1##
Where tested in this assay, the compounds of the invention gave the
following results, showing them to exhibit a moderate to very
significant inhibition of IL 1-induced protease secretion:
______________________________________ Compound of Dose %
Inhibition Example No. (.mu.M) (I.S.D)
______________________________________ 1A 10 35 .+-. 11 1B 10 98
.+-. 1 5 93 .+-. 5 1 94 0.1 67 2A 10 34 .+-. 26 2B 10 85 .+-. 6 1
36 3A 10 .ltoreq.20 3B 10 81 .+-. 2 1 27 4 10 43 5 46 1 38 5 10 79
.+-. 14 5 55 .+-. 22 1 37 .+-. 0 6 10 32 5 19 7 10 25 5 26 1 10 8
10 16 5 37 1 3 9 10 58 5 53 1 30 10 10 96 1 91 0.1 70 11 10 88 1 50
0.1 58 12 10 68 1 61 0.1 57
______________________________________
* * * * *